Research Doxycycline And Adriamycin On BGC823Apoptosis Of Gastric Cancer Cells And Its Mechanism Of Action

ObjectiveThrough the use of doxycycline, doxorubicin monotherapy and two drugcombination group effect on gastric cancer BGC823cells, to observe the effectof three drugs on the apoptosis of gastric cancer BGC823cells, analysis ofBcl-2, Bax, Caspase-3, expression of Notch-1protein in inducing tumor cellsapoptosis, and to explore its mechanism, the therapy period to provide efficientand low toxicity for clinical tumor chemotherapy.MethodsThe gastric cancer BGC823cell culture and intervention, respectively to afinal concentration of5,10,15,20,25ug/ml2,2.5, doxycycline;3,3.5,4ug/mladriamycin; combined treatment group effect of5/2.0,5/2.5,5/3.0,5/3.5,5/4.0ug/ml in human gastric cancer cells BCG823, color four methyl thiazolyltetrazolium (MTT) than the inhibition of gastric cancer cell was detected drug.The high concentration of drug taking and divided into4groups: A: controlgroup; doxycycline group B:5ug/ml; Adriamycin group C:4.0ug/ml; thecombination group5/4.0ug/ml. By Wright Gimsa staining staining, the morphological changes of the cells were observed under a microscope; flowcytometry was used to detect effects of tumor cell apoptosis rate; the changesof Western blot method to detect Bcl-2, Bax, Caspase-3, Notch-1proteinexpression.Results1.With doxycycline, adriamycin and combination group, BGC823cells weretreated with24h,48h and72h, MTT results show: the inhibitory effect ofdifferent drugs on the proliferation of BGC823cells were, in a time dosedependent manner, with the increase of drug concentration and the extension ofthe time, on gastric cancer cell line BGC823proliferation effect of the moreobvious.2.By Wright Gimsa staining staining, the morphological changes of the cellswere observed under a microscope, the control group of gastric cancer BGC823cells were polygon or oval, clear cytoplasm, monolayer growth, by all threegroups after drug treatment, the survival number of cells and cytoplasmictransparency decreased significantly, cell membrane integrity but the foamingphenomenon.3.Flow cytometry showed, compared with the control group, adriamycinand combination group on BGC823cell, plays a significant role of apoptosisinduced. When the combined use of doxycycline and doxorubicin, significantlyenhanced the apoptosis induced by adriamycin.4.The Western results by computer image analysis, Western Blot testresults show that, compared with the control group, adriamycin group, Baxincreased and Bcl-2decreased, the combination group Bax increased and Bcl-2decreased more obviously. Compared with the control group, adriamycin singledrug group Notch1, caspase-3in the cells increased, expression of Notch1,caspase-3protein in the combination group increased significantly.Conclusions1.Doxycycline and adriamycin combination is better than doxycycline anddoxorubicin monotherapy group. 2.Compared with doxorubicin monotherapy, combination therapy canpromote apoptosis in BGC823human gastric cancer cells, expression, whichmay be related to the up regulation of Notch-1activation of Caspase-3,activation of Bax and suppression of Bcl-2.3.The combined application of doxycycline as apoptosis apoptosispromoter and chemotherapeutic drugs can inhibit tumor cell.